The present invention provides a composition capable of maintaining antioxidative activity in blood high by keeping the reduced coenzyme Q10 ratio in blood high after taking coenzyme Q10 even in aged mammals.

The present invention relates to a composition which comprises reduced coenzyme Q10 as an active component and enhances the antioxidative activity in blood of aged mammals.

13. A method for administering a composition to aged mammals, wherein said composition comprises reduced coenzyme Q10 represented by the following formula (1) as an active component

14. The method according to claim 13, which enhances the antioxidative activity in blood of aged mammals.

15. The method according to claim 13, which is for treating or preventing diseases related to oxidative stress.

16. The method according to claim 13, which is for treating or preventing diseases related to intravascular oxidative stress.

17. The method according to claim 13, wherein said composition further comprises oxidized coenzyme Q10 represented by the following formula (2)

18. The method according to claim 13, wherein said composition further comprises an antioxidant.

19. The method according to claim 14, wherein said composition further comprises oxidized coenzyme Qlo represented by the following formula (2).

20. The method according to claim 15, wherein said composition further comprises oxidized coenzyme Qlo represented by the following formula (2).

21. The method according to claim 16, wherein said composition further comprises oxidized coenzyme Qlo represented by the following formula (2).

22. The method according to claim 14, wherein said composition further comprises an antioxidant.

23. The method according to claim 15, wherein said composition further comprises an antioxidant.

24. The method according to claim 16, wherein said composition further comprises an antioxidant.

Description:

TECHNICAL FIELD

The present invention relates to a composition which comprises reduced coenzyme Q10 as an active component and enhances the antioxidative activity in blood of aged mammals.

BACKGROUND ART

Active oxygen contributes to physiologically important metabolism and plays important roles in vital activities on some occasions by, for example, producing biologically active components, killing pathogenic bacteria, and playing an important role involving anticancer action. On the other hand, excess generation of active oxygen, which cannot be controlled, is considered to cause some damage to the living body via denaturation of nucleic acids and/or proteins, or a peroxidation reaction of lipids. The condition wherein the amount of active oxygen is not controlled and excess amount of active oxygen occurs is called oxidative stress condition. In recent years, it has become clear that oxidative stress causes some adverse effect on various diseases.

As the diseases related to oxidative stress, there has been reported arteriosclerosis, cancer, cerebral ischemia, hepatic dysfunction, diabetes mellitus, neurological disorder, renal disorder, hepatic cirrhosis, arthritis, retinopathy of prematurity, ocular uveitis, retinal siderosis, senile cataract, dysfunction due to an adverse effect derived from radiation therapy, asbestosis, bronchial injury due to smoking, dysfunction due to an adverse effect derived from carcinostatic drug use, cerebral edema, pulmonary edema, pedal edema, cerebral infarction, hemolytic anemia, progeria, epilepsy, Alzheimer's disease, Down's syndrome, Parkinson's disease, Behcet's disease, Crohn's disease, Kawasaki disease, Weber-Christian disease, collagen disease, progressive systemic sclerosis, dermatitis herpetiformis, immunologic deficiency syndrome, and the like diseases. To such diseases, availability of antioxidants (radical scavengers) which are considered to have oxidative stress-reducing effect has been examined for years, and anti-inflammatory drugs showing a radical scavenging effect have been developed. In addition, according to the fact that a novel substance (Radicut) showing an antioxidative effect was recently certified as a pharmaceutical preparation, availability of the antioxidants in reducing oxidative stress can be said obvious.

Coenzyme Q10 is a component widely distributed in living organisms, and performs important function as a component of the electron transport system of mitochondria, which is one of organelle. Also, coenzyme Q10 was found to have another important effect, namely the antioxidative effect, and has drawn attention. Having the antioxidative activity, coenzyme Q10 is expected to have prevention and improvement effects on the above-mentioned diseases which are supposedly related to oxidative stress, and has been increasingly taken as a supplement in recent years.

Coenzyme Q10 occurs as its oxidized form or reduced form. One showing the antioxidative activity is reduced coenzyme Q10 and therefore no antioxidative activity is exerted even when there are enormous amount of oxidized forms (Non-Patent Document 1 and 2). Accordingly, at the time of oxidized coenzyme Q10 intake, oxidized coenzyme Q10 is necessary to be converted into its reduced form in the living body for showing its antioxidative activity. That is, for enhancing the antioxidative activity and reducing oxidative stress in the living body by taking coenzyme Q10, it is very important that coenzyme Q10 occurs as its reduced form in the organs and blood of the living body, where coenzyme Q10 is transported via blood.

Generally, it is said that oxidized coenzyme Q10 orally taken as a supplement is absorbed into the small intestine, enzymatically reduced to reduced coenzyme Q10 with consuming reducing equivalent of reduced nicotineamide adenine dinucleotide phosphate (NADPH) and the like as a substrate, and then released into blood in the form integrated in lipoproteins (Non-Patent Document 1 and 3). That is, oxidized coenzyme Q10 intake also increases reduced coenzyme Q10 in the living body. On the other hand, an oxidized form and reduced form of coenzyme Q10 coexist in plasma. In recent years, it has become clear that the reduced coenzyme Q10 ratio in plasma is useful as an oxidative stress marker. This is attributed to the fact that reduced coenzyme Q10 which has been once oxidized is not reduced again in plasma since no enzyme capable of reducing oxidized coenzyme Q10 exists in blood. It has been reported that, as for liver diseases known to be exacerbated due to oxidative stress, the oxidized coenzyme Q10 ratio increases depending on the severity of the disease (Non-Patent Document 4). In this manner, the effect of oxidative stress exposure is shown in the reduced coenzyme Q10 ratio in plasma, and thus it is supposed that increased ratio of reduced coenzyme Q10 in plasma results from an enhanced antioxidative activity in the living body.

Non-Patent Document 5 discloses that, when a 35-year-old healthy person took oxidized coenzyme Q10, the ratio of reduced coenzyme Q10 to oxidized coenzyme Q10 was unchanged despite the amount of reduced coenzyme Q10 in plasma increased. That is, although oxidized coenzyme Q10 is reduced and converted into its reduced form during adsorption, the antioxidative activity in the living body cannot be enhanced and the ratio of reduced coenzyme Q10 to the total amount of coenzymes in plasma cannot increase. No composition showing such activity has been known.

Furthermore, for aged humans and aged mammals considered to be exposed to oxidative stress for years and have lowered antioxidative activity, no method nor composition for increasing the reduced coenzyme Q10 ratio to the total amount of coenzymes in plasma has also been unknown.

SUMMARY OF THE INVENTION

Accordingly, the present invention has for its object to provide a composition capable of maintaining antioxidative activity in blood high, even in aged mammals, by keeping the reduced coenzyme Q10 ratio in blood high after coenzyme Q10 intake.

The present inventors have made intensive investigations to solve the above-mentioned subjects and, as a result, they surprisingly found that the reduced coenzyme Q10 ratio in blood can be kept high even in aged mammals by the intake of a composition comprising reduced coenzyme Q10. Such and other findings have led to completion of the present invention.

Thus, the present invention relates to

a composition

which comprises reduced coenzyme Q10 represented by the following formula (1), as an active component.

Also, the present invention relates to

a method for producing a composition

which comprises mixing reduced coenzyme Q10 with a carrier acceptable as a pharmaceutical preparation or food.

The above-mentioned composition is preferable for administration to aged mammals, and is also preferable for enhancing the antioxidative activity in blood of aged mammals.

Furthermore, the present invention relates to

a method for enhancing the antioxidative activity in blood

which comprises administering reduced coenzyme Q10.

The above method is preferably used for aged mammals.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is now described in detail.

The composition of the invention which comprises reduced coenzyme Q10 as an active component and enhances the antioxidative activity in blood of aged mammals is described.

Reduced coenzyme Q10 according to the invention is represented by the above formula (1).

The method for obtaining reduced coenzyme Q10 is not particularly restricted and employable as such methods are, for example, a method comprising obtaining coenzyme Q10in the conventional manner such as synthesis, fermentation or extraction from natural sources, and then concentrating a reduced coenzyme Q10 fraction in the effluent by chromatography. Also employable for obtaining reduced coenzyme Q10 is a method comprising reacting an existing highly pure oxidized coenzyme Q10 with an ordinary reducing agent such as sodium borohydride or sodium dithionite (sodium hydrosulfite). Further, a strain containing reduced coenzyme Q10, and the like can also be used.

In the invention, the composition which comprises reduced coenzyme Q10 as an active component may further comprise oxidized coenzyme Q10 represented by the following formula (2).

In this case, the content of reduced coenzyme Q10 to the total coenzyme Q10 is not particularly restricted, but the lower limit thereof is preferably not lower than 80% by weight, more preferably not lower than 90% by weight, still more preferably not lower than 95% by weight, and particularly preferably not lower than 98% by weight. The upper limit of reduced coenzyme Q10 is preferably not higher than 100% by weight.

The ratio of the reduced form to the total coenzyme Q10 is generally determined by a method comprising quantifying oxidized coenzyme Q10 and reduced coenzyme Q10 in a sample by high performance liquid chromatography (HPLC) using an UV detector and calculating the ratio from the volume ratio obtained, or a method comprising using an HPLC system incorporated with an electrochemical detector for calculating the ratio of oxidized coenzyme Q10 and reduced coenzyme Q10 from the peak area. The HPLC system incorporated with an electrochemical detector makes it possible to specifically determine oxidation-reduction substances and has high sensitivity, and thus is highly useful for determining trace amount of reduced coenzyme Q10 in the living body or samples. In the present invention, all the reduced coenzyme Q10 ratios are determined by the HPLC system incorporated with an electrochemical detector.

The content of coenzyme Q10 in the composition of the invention is preferably 0.001 to 99% by weight, and more preferably 0.01 to 20% by weight.

The aged mammals cited in the invention refer to, as for human, individuals of 40 years old or older. The use to humans of 60 years old or older is particularly preferable. As for rats, individuals of 50 weeks old or older are referred to. And as for other animals, individuals equivalent to humans of 40 years old or older are referred to.

The composition of the invention can be used in pharmaceutical products, functional foods, food materials, or the like. The functional foods cited herein refer to products for maintaining or improving health by oral intake of products other than pharmaceutical products, such as oral supplements, foods for specified health uses, health foods, or dietary supplements.

In producing the composition of the invention, the content, dosage form, preservation method, and preservation form of reduced coenzyme Q10 can be arbitrary determined according to the application, such as use in pharmaceutical products, health foods or foods. The composition of the present invention is produced by mixing reduced coenzyme Q10 with a carrier acceptable as a pharmaceutical preparation or food and, according to need, oxidized coenzyme Q10 and/or an antioxidant. The carrier acceptable as a pharmaceutical preparation or food is not particularly restricted and there may be mentioned, for example, an excipient, a disintegrant, a lubricant, a binder, a colorant, a coagulation inhibitor, an absorption promoter, a solubilizing agent, a stabilizer, and the like.

The dosage form of the composition of the invention is not particularly restricted and may be, for example, solution preparations, powders, granules producible by adding a binder, powders coated with a coating agent, and capsule preparations producible by filling powders, granules or coated powders into capsules. The solution preparations are not particularly restricted and there may be mentioned, for example, a drink, an injection, an infusion, and the like. Also employable are soft capsule preparations producible by adding natural oil, higher fatty acid oil, a higher fatty acid monoglyceride, a surfactant, or mixture thereof, etc. and filling that in its oily state into capsules. In this case, as the capsule, gelatin-based ones or ones based on other soluble polymer substances than gelatin can be used, for example. Also, microcapsules are included in such capsules.

EFFECT OF THE INVENTION

According to the present invention, a composition capable of keeping the reduced coenzyme Q10 ratio in blood high, even in aged mammals, by coenzyme Q10 intake, and therefore capable of maintaining antioxidative activity in blood high is provided.

BEST MODE FOR CARRYING OUT THE INVENTION

The following Examples and Preparation Examples illustrate the present invention in further detail. These Examples and Preparation Examples are, however, by no means limitative of the scope of the invention.

In these Examples, the ratio of reduced coenzyme Q10 to the total coenzyme Q10 was determined using the HPLC system incorporated with an electrochemical detector (product of Shiseido Co., Ltd.) and a reduction column (product of Shiseido Co., Ltd.).

REFERENCE EXAMPLE 1

10 SD rats (5-week-old, males) were divided into 2 groups each consisted of 5 rats so that each group had the same average body weight value. To one group, a solution of reduced coenzyme Q10 in soybean oil (containing about 1% of oxidized coenzyme Q10) was orally administered, and to the other group, a solution of oxidized coenzyme Q10 in soybean oil was orally administered, each at the dose of 100 mg/kg body weight in coenzyme Q10 equivalent. Then, the reduced coenzyme Q10 ratio in plasma was determined with time lapses. Blood was collected after 1, 2, 4 and 8 hours of oral administration, and centrifuged to obtain plasma. The concentration of reduced coenzyme Q10 and oxidized coenzyme Q10 in the obtained plasma was quantified using HPLC in order to determine the reduced coenzyme Q10 ratio.

FIG. 1 shows the change of the ratio of the reduced form of coenzyme Q10 in plasma. It shows that there was almost no difference in the reduced coenzyme Q10 ratio after 2 hours or later of administration between the administration of reduced coenzyme Q10 and that of oxidized coenzyme Q10.

EXAMPLE 1

10 SD rats (54-week-old, males) were divided into 2 groups each consisted of 5 rats so that each group had the same average body weight value. To one group, a solution of reduced coenzyme Q10 in soybean oil (containing about 1% of oxidized coenzyme Q10) was orally administered, and to the other group, a solution of oxidized coenzyme Q10 in soybean oil was orally administered, each at the dose of 100 mg/kg body weight in coenzyme Q10 equivalent. Then, the reduced coenzyme Q10 ratio in plasma was determined with time lapses. Blood was collected after 1, 2, 4 and 8 hours of oral administration, and centrifuged to obtain plasma. The concentration of reduced coenzyme Q10 and oxidized coenzyme Q10 in the obtained plasma was quantified using HPLC in order to determine the ratio of the reduced form.

FIG. 2 shows the change of the ratio of the reduced form of coenzyme Q10 in plasma. It shows that, when reduced coenzyme Q10 was administered, the ratio of the reduced form remained around 90%. Comparatively, when oxidized coenzyme Q10 was administered, the ratio of the reduced form in plasma was about 76% after 1 hour of administration. The ratio had a tendency to increase with the lapse of time but, even after 8 hours of administration, the ratio remained about 87%, and did not increase to 90%.

EXAMPLE 2

10 SD rats (62-week-old, males) were divided into 2 groups each consisted of 5 rats so that each group had the same average body weight value. To one group, a solution of reduced coenzyme Q10in soybean oil (containing about 1% of oxidized coenzyme Q10) was orally administered, and to the other group, a solution of oxidized coenzyme Q10 in soybean oil was orally administered, each at the dose of 100 mg/kg body weight in coenzyme Q10 equivalent. Then, the reduced coenzyme Q10 ratio in plasma was determined with time lapses. Blood was collected after 1, 2, 4 and 8 hours of oral administration, and centrifuged to obtain plasma. The concentration of reduced coenzyme Q10 and oxidized coenzyme Q10 in the obtained plasma was quantified using HPLC in order to determine the ratio of the reduced form.

FIG. 3 shows the change of the ratio of the reduced form of coenzyme Q10 in plasma. It shows that, when reduced coenzyme Q10 was administered, the ratio of the reduced form remained around 90%. Comparatively, when oxidized coenzyme Q10 was administered, the ratio of the reduced form in plasma was about 74% after 1 hour of administration. The ratio had a tendency to slightly increase with the lapse of time but, even after 8 hours of administration, the ratio remained about 78%.

PREPARATION EXAMPLE 1

Olive oil was warmed to 60° C., and reduced coenzyme Q10 (containing about 1% of oxidized coenzyme Q10) melted at 60° C. was added for dissolution. Thereto, vitamin E was added little by little for attaining a uniform mixture, and soft capsules were prepared in the conventional manner. Soft capsule preparations containing 20 mg of reduced coenzyme Q10 in each capsule were obtained.

Reduced coenzyme Q10

20 parts by weight

(containing about 1% of oxidized coenzyme Q10)

Vitamin E

15 parts by weight

Olive oil

350 parts by weight

PREPARATION EXAMPLE 2

Reduced coenzyme Q10 (containing about 1% of oxidized coenzyme Q10) was dissolved in acetone, and then allowed to be adsorbed on crystalline cellulose (fine powders) in order to be dried. The obtained product was mixed with corn starch to give powder preparations in the conventional manner.

Reduced coenzyme Q10

10 parts by weight

(containing about 1% of oxidized coenzyme Q10)

Crystalline cellulose (fine powders)

40 parts by weight

Corn starch

55 parts by weight

PREPARATION EXAMPLE 3

Reduced coenzyme Q10 (containing about 1% of oxidized coenzyme Q10) was dissolved in acetone, and then allowed to be adsorbed on crystalline cellulose (fine powders) in order to be dried. The obtained product was mixed with corn starch, lactose, carboxymethyl cellulose, and magnesium stearate. Then, an aqueous solution of polyvinylpyrrolidone was added thereto as a binder and the mixture was granulated in the conventional manner to obtain powders. With these powders, talc was mixed as a lubricant, and tablets containing 20 mg of reduced coenzyme Q10in each tablet were obtained.

Reduced coenzyme Q10

20 parts by weight

(containing about 1% of oxidized coenzyme Q10)

Corn starch

25 parts by weight

Lactose

15 parts by weight

Calcium carboxymethyl cellulose

10 parts by weight

Crystalline cellulose (fine powders)

40 parts by weight

Polyvinylpyrrolidone

5 parts by weight

Magnesium stearate

3 parts by weight

Talc

10 parts by weight

PREPARATION EXAMPLE 4

The following ingredients were granulated in the conventional manner and filled into gelatin hard capsules. Capsule preparations containing 20 mg of reduced coenzyme Q10 in each capsule were obtained.

Reduced coenzyme Q10

20 parts by weight

(containing about 1% of oxidized coenzyme Q10)

Crystalline cellulose (fine powders)

40 parts by weight

Corn starch

20 parts by weight

Lactose

62 parts by weight

Magnesium stearate

2 parts by weight

Polyvinylpyrrolidone

3 parts by weight

PREPARATION EXAMPLE 5

Rapeseed oil was warmed to 60° C., and reduced coenzyme Q10 (containing about 1% of oxidized coenzyme Q10) melted at 60° C. was added for dissolution. Thereto, beeswax was added as a stabilizer and soft capsules were prepared in the conventional manner. Soft capsule preparations containing 50 mg of reduced coenzyme Q10in each capsule were obtained.

Reduced coenzyme Q10

50 parts by weight

(containing about 1% of oxidized coenzyme Q10)

Rapeseed oil

300 parts by weight

Beeswax

40 parts by weight

COMPARATIVE PREPARATION EXAMPLE 1

Soft capsule preparations containing oxidized coenzyme Q10 were obtained in the same manner as Preparation Example 5 except that oxidized coenzyme Q10 was used in lieu of reduced coenzyme Q10.

Oxidized coenzyme Q10

50 parts by weight

Rapeseed oil

300 parts by weight

Beeswax

40 parts by weight

EXAMPLE 3

Two soft capsules obtained in Preparation Example 5 and Comparative Preparation Example 1 (containing 100 mg of coenzyme Q10) were fed to two groups of healthy aged people (20 male and female individuals in each group, age: 64 to 77 years old), and the effect on the reduced coenzyme Q10 ratio in plasma was evaluated. The results are shown in Table 1.

TABLE 1

Change of reduced coenzyme Q10 ratio

(%) in plasma

After 2 hours of

After 6 hours of

feeding

feeding

Group fed with

−0.86

−0.09

oxidized coenzyme Q10

Group fed with

0.05

0.51

reduced coenzyme Q10

In the group fed with oxidized coenzyme Q10, the ratio of reduced coenzyme Q10 to the total coenzyme Q10 in plasma rather decreased after 2 hours of feeding, and the ratio recovered to almost the same level as before feeding after 6 hours of feeding. This result is nearly identical to the result shown in Non-Patent Document 5. It is confirmed that feeding of oxidized coenzyme Q10 rather decreases the ratio of the reduced form in plasma. It is supposedly resulted from lowered antioxidative activity due to consumption of the reducing equivalent in bodies for reducing the oxidized form. On the contrary, in the group fed with reduced coenzyme Q10, the ratio of reduced coenzyme Q10 to the total coenzyme Q10 in plasma increased with the lapse of time since 2 hours after administration, which shows the antioxidative activity in bodies increased.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a line graph showing the time course of reduced coenzyme Q10 ratio in plasma of 5-week-old SD rats according to Reference Example 1. The ordinate shows the ratio (%) of the reduced form to the total coenzyme Q10 in plasma. The abscissa shows the lapse of time after administration of coenzyme Q10. The symbols shows as follows; : the group fed with reduced coenzyme Q10, and □: the group fed with oxidized coenzyme Q10. The values plotted on the graph are the average value of 5 rats in each group.

FIG. 2 is a line graph showing the time course of reduced coenzyme Q10 ratio in plasma of 54-week-old SD rats according to Example 1. The ordinate shows the ratio (%) of the reduced form to the total coenzyme Q10 in plasma. The abscissa shows the lapse of time after administration of coenzyme Q10. The symbols shows as follows; : the group fed with reduced coenzyme Q10, and □: the group fed with oxidized coenzyme Q10. The values plotted on the graph are the average value of 5 rats in each group.

FIG. 3 is a line graph showing the time course of reduced coenzyme Q10 ratio in plasma of 62-week-old SD rats according to Example 2. The ordinate shows the ratio (%) of the reduced form to the total coenzyme Q10 in plasma. The abscissa shows the lapse of time after administration of coenzyme Q10. The symbols shows as follows; : the group fed with reduced coenzyme Q10, and H: the group fed with oxidized coenzyme Q10. The values plotted on the graph are the average value of 5 rats in each group.